Glucose Uptake-Glo™ Assay

Measuring Glucose Uptake

Glucose metabolism is a key process in many organisms. A lack of insulin-stimulated glucose uptake is associated with type 2 diabetes, while high glucose uptake is a sign of the high glycolytic rates associated with cancer. Measuring glucose uptake can determine the effects of various treatments for diabetes and cancer.

The gold standard method of assaying glucose uptake relies on detection of radio-labeled glucose analogs. Alternative colorimetric and fluorometric glucose uptake assays often lack the sensitivity and robustness needed to reliably measure glucose uptake in cells.

A Simple, Non-Radioactive Glucose Uptake Assay

The Glucose Uptake-Glo™ Assay is a plate-based, homogeneous bioluminescent method for measuring glucose uptake in cells, based on the detection of 2-deoxyglucose-6-phosphate (2DG6P).

Simple, homogeneous protocol: After addition of 2DG, there are no wash steps—all steps are additions.
Sensitive with broad linearity: The Glucose Uptake-Glo™ Assay can detect 0.5–30µM 2DG6P, and generates a signal-to-background > 3 with as few as 5,000 cells.
Compatible with automation: The “add-mix-measure” format is compatible with automated and high-throughput workflow; reactions are scalable for use in 96- and 384-well plates.
Reliable and reproducible: The Glucose Uptake-Glo™ Assay yields Z factors > 0.5

"Promega products are reliable and can be purchased without any hesitation. My experiment went very well as expected and we have more plans to conduct uptake experiments using the Glucose Uptake-Glo™ Assay."

Dr. Vivek Pandey, Postdoctoral Scholar, University of Kentucky.

A comparison of the Glucose Uptake-Glo™ Assay with the conventional radioactive method

2-deoxyglucose (2DG) is transported into cells and phosphorylated to produce 2-deoxyglucose-6-phosphate (2DG6P). The addition of Stop Buffer stops 2DG transport, lyses cells, destroys any NADPH within the cells and inactivates proteins. The addition of Neutralization Buffer neutralizes the solution before addition of the 2DG6P Detection Reagent. The glucose-6-phosphate dehydrogenase (G6PDH) within the reagent oxidizes 2DG6P to 6-phosphodeoxygluconate (6PDG) and reduces NADP+ to NADPH. The reductase uses the NADPH to convert the proluciferin to luciferin, which is then used by luciferase to produce light.

Measure Metabolic Changes in Cancer and Adipocyte Cell Lines

MCF7 Cancer Cells

In the cancer model, when cells are oxygen-starved, the hypoxic conditions shift cellular metabolism from oxidative phosphorylation to glycolysis. This results in increased glucose uptake.

MCF7 cells grown under hypoxia (1% oxygen) show an increase in Glucose Uptake-Glo™, indicating an increased glycolytic rate. The same cells demonstrate no significant change in viability using the RealTime-Glo™ and CellTiter-Fluor™ Assays.

Adipocyte Cells

Using the adipocyte model, we demonstrate that changes in glucose uptake for 3T3L1-MBX adipocytes did not have significant effects on cell health. Multiplexing the RealTime-Glo™ Assay with the Glucose Uptake-Glo™ Assay separates immediate effects on glucose uptake from global effects on cell health.

13506MB-W-a — Results of the Glucose Uptake-Glo™ Assay when adipocyte cells are treated under various conditions, where "None" indicates the basal level of glucose uptake. Insulin induces translocation of glucose transporters to the cell surface, and thus increases glucose uptake above basal levels. Cytochalasin B is a glucose transporter inhibitor which decreases glucose uptake. LY294002, a phosphatidylinositol 3-kinase (PI3K) inhibitor, is an essential insulin signaling enzyme which decreases glucose uptake relative to insulin alone.

13506MB-W-b — Results of the RealTime-Glo™ Assay when adipocyte cells are treated under various conditions, where "None" indicates the basal level of glucose uptake. Although there are dramatic changes in glucose uptake, as seen in the accompanying panel, there are minimal changes in viability as measured by the RealTime-Glo™ Assay.

Protocolos

Complete Protocol

Glucose Uptake-Glo™ Assay Technical ManualPDF (2409 KB) – English

Contenido

Item	Part #	Presentación
Luciferase Reagent	J151A	1 × 5ml
Stop Buffer	J152A	1 × 15ml
Neutralization Buffer	J153A	1 × 15ml
NADP+	J154A	1 × 50μl
G6PDH	J155A	1 × 125μl
2DG	J156A	1 × 250μl
2DG6P Standard	J157A	1 × 50μl
Reductase	G884C	1 × 25μl
Reductase Substrate	G885A	1 × 55μl

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

Patentes y Exclusiones

U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.

Contenido

Item	Part #	Presentación	Concentración
Luciferase Reagent	J151B	1 × 10ml
Stop Buffer	J152A	1 × 15ml
Neutralization Buffer	J153A	1 × 15ml
NADP+	J154B	1 × 100μl
G6PDH	J155B	1 × 250μl
2DG	J156A	1 × 250μl
2DG6P Standard	J157A	1 × 50μl
Reductase	G884A	1 × 55μl	6mg/ml
Reductase Substrate	G885A	1 × 55μl

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

Patentes y Exclusiones

U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.

Contenido

Item	Part #	Presentación	Concentración
Luciferase Reagent	J151C	1 × 50ml
Stop Buffer	J152A	1 × 15ml
Neutralization Buffer	J153A	1 × 15ml
NADP+	J154C	1 × 500μl
G6PDH	J155C	1 × 1.25ml
2DG	J156A	1 × 250μl
2DG6P Standard	J157A	1 × 50μl
Reductase	G884B	1 × 275μl	6mg/ml
Reductase Substrate	G885A	1 × 55μl

SDS

Search for SDS

Certificado de Análisis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Condiciones de Almacenaje

Patentes y Exclusiones

U.S. Pat. Nos. 9,273,343 and 9,951,372, European Pat. No. 2751089, Japanese Pat. No. 6067019 and other patents pending.

Recursos

Artículos

High-pressure oxygen rewires glucose metabolism of patient-derived glioblastoma cells and fuels inflammasome response

A Simple, Non-Radioactive Assay to Measure Glucose Uptake Inside Cells

Measuring Glucose Uptake

A Simple, Non-Radioactive Glucose Uptake Assay

A comparison of the Glucose Uptake-Glo™ Assay with the conventional radioactive method